Helicobacter felis Paster et al.
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
2. Aseptically transfer the thawed suspension into a #260 broth tube. This broth can now be used to inoculate an agar slant(s), plate(s), additional broth tube(s), or the preferred biphasic culture.
3. To obtain a biphasic culture, add 0.5 ml of the suspension to a #260 slant. The resulting pool at the bottom of the slant is where the best, most rapid growth will occur.
4. Incubate at 37oC under microaerophilic conditions using an anaerobe jar with an active catalyst and a microaerophilic gas generator pack, or other acceptable method, to obtain microaerophilic conditions. Incubate slant with cap loosen.
Growth on agar takes longer than the biphasic culture. Colonies are flat with some spreading. It is essential to use fresh, moist plates. The cells do not Gram stain well using traditional procedures. For best results, use a basic fuchsin counterstain in place of the safranin.
Once good growth in obtained, transfer or freeze the culture. Adding an equal amount of 20% sterile glycerol to pooled broth from several biphasic slants, followed by freezing in liquid nitrogen or ultra-low temperature freezer is recommended.
See also Helicobacter species, General Procedures in ATCC Bacteria and Bacteriophages, 19th edition, 1996, p.471.
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Paster BJ, et al. Phylogeny of Helicobacter felis sp. nov., Helicobacter mustelae, and related bacteria. Int. J. Syst. Bacteriol. 41: 31-38, 1991. PubMed: 1704791